猪细小病毒重组酶聚合酶扩增快速检测方法的建立

doi:10.16431/j.cnki.1671-7236.2017.11.030

Abstract

Abstract:

This study was aimed to develop a simple and rapid method for the porcine parvovirus (PPV) with recombinase polymerase amplification (RPA), which could be a novel and reliable tool for the control and detect of PPV. The RPA was developed using specific primers for the conserved region of PPV VP2 gene. The reaction time was optimized,the RPA reaction could amplify the PPV, and was performed successfully at 38℃ for 30 min in a water bath. The results showed that the detection limit of RPA was 102 copies of plasmid DNA, which was the same as the Real-time quantitative PCR applied in this study, and 100 times more sensitive than conventional PCR. For the clinical samples from the suspected PPV-infected pigs, the positive detection ratio was 82.6% for RPA, which was lower than that of Real-time quantitative PCR (86.9%), but was much higher than conventional PCR (66.7%). The PPV RPA assay developed in the study was simple, rapid and reliable, and was suitable for rapid detection of PPV.

Key words: porcine parvovirus (PPV); isothermal amplification; recombinase polymerase amplification (RPA); rapid detection

CLC Number:

S852.65⁺9.2

LIU Li-bing, WANG Jian-chang, JING Mei, WANG Jin-feng, YUAN Wan-zhe. Development of the Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Parvovirus[J]. , 2017, 44(11): 3320-3326.

References

[1] Molitor T W, Joo H S, Collett M S. Porcine parvovirus:Virus purification and structural and antigenic properties of virion polypeptides[J]. Journal of Virology, 1983, 45(2):842-854.
[2] Mayr A, Bachmann P A, Siegl G, et al. Characterization of a small porcine DNA virus[J]. Arch Gesamte Virusforsch,1968,25(1):38-51.
[3] Ren X, Tao Y, Cui J, et al. Phylogeny and evolution of porcine parvovirus[J].Virus Research, 2013, 178(2):392-397.
[4] Rueda P,Fominaya J, Langeveld J P M, et al. Effect of different baculovirus inactivation procedures on the integrity and immunogenicity of porcine parvovirus-like particles[J]. Vaccine, 2000, 19(7-8):726-734.
[5] Kim J, Chung H K, Chae C. Association of porcine circovirus 2 with porcine respiratory disease complex[J]. Veterinary Journal, 2003, 166(3):251-256.
[6] Ellis J A, Bratanich A, Clark E G, et al. Coinfection by porcine circoviruses and porcine parvovirus in pigs with naturally acquired postweaning multisystemic wasting syndrome[J]. Journal of Veterinary Diagnostic Investigation, 2000, 12(1):21-27.
[7] Lager K M, Mengeling W L. Porcine parvovirus associated with cutaneous lesions in piglets[J].Journal of Veterinary Diagnostic Investigation, 1994, 6(3):357-359.
[8] Chen H Y, Li X K, Cui B A, et al. A TaqMan-based Real-time polymerase chain reaction for the detection of porcine parvovirus[J]. Journal of Virological Methods, 2008, 156(1-2):84-88.
[9] Piepenburg O, Williams C H, Stemple D L, et al. DNA detection using recombination proteins[J]. PLoS Biology, 2006, 4(7):e204.
[10] 付朋飞,乔涵,张宇,等.表达猪细小病毒VP2蛋白的猪伪狂犬病病毒rPRV-VP2株的生物学特性[J].中国兽医学报,2017,37(1):11-17.
[11] 王林青,付朋飞,乔涵,等.表达猪细小病毒VP2蛋白和猪IL-18重组伪狂犬病毒的构建[J].中国兽医学报,2015, 35(9):1422-1428.
[12] Cao L, Chen J, Wei Y, et al. Porcine parvovirus induces activation of NF-κB signaling pathways in PK-15 cells mediated by Toll-like receptors[J]. Molecular Immunology, 2017, 85:248.
[13] Fernandes S, Boisvert M, Tijssen P. Genetic elements in the VP region of porcine parvovirus are critical to replication efficiency in cell culture[J]. J Virol,2011, 85(6):3025-3029.
[14] Xu Y G, Cui L C, Wang H W, et al. Characterization of the capsid protein VP2 gene of a virulent strain NE/09 of porcine parvovirus isolated in China[J]. Research in Veterinary Science, 2013, 94(2):219-224.
[15] Ji P, Liu Y, Chen Y, et al. Porcine parvovirus capsid protein expressed in Escherichia coli, self-assembles into virus-like particles with high immunogenicity in mice and guinea pigs[J]. Antiviral Research, 2017, 139:146.
[16] 中华人民共和国国家质量监督检验检疫总局. 猪细小病毒病聚合酶链反应操作规程(SN/T 1874-2007)[S].北京:中国标准出版社,2007.
[17] Wilhelm S, Zimmermann P, Selbitz H J, et al. Real-time PCR protocol for the detection of porcine parvovirus in field samples[J].Journal of Virological Methods, 2006, 134(1-2):257-260.
[18] Yun J J, Heisler L E, Hwang I I, et al. Genomic DNA functions as a universal external standard in quantitative Real-time PCR[J]. Nucleic Acids Research, 2006, 34(12):e85.
[19] 崔宇超,崔尚金. 猪细小病毒的流行语防治进展[J].养猪,2015,1:123-128.
[20] 邹敏,李陆梅,宫晓,等. 猪细小病毒PCR检测方法的建立与应用[J].中国动物检疫,2016,33(8):94-97.
[21] 沈志强,王金良,郭显坡,等. SYBR Green Ⅰ实时荧光定量PCR检测猪细小病毒方法建立及初步应用[J].中国兽医学报,2011, 31(1):11-15.
[22] 陈星瑶,张子群,王晓龙. 猪细小病毒LAMP检测方法的建立[J].中国动物保健,2014,16(8):9-12.
[23] 崔宇超,彭永刚,马兴杰,等. 猪细小病毒高效纳米PCR检测方法的建立[J].中国预防兽医学报,2014, 36(4):286-288.
[24] Mckillen J, Hjertner B, Millar A, et al. Molecular beacon Real-time PCR detection of swine viruses[J].Journal of Virological Methods, 2007, 140(1-2):155.
[25] Yu H Q, Cai X Q, Lin Z X, et al. Rapid and specific detection of porcine parvovirus using Real-time PCR and high resolution melting (HRM) analysis[J]. BMC Veterinary Research, 2015, 11(1):1-5.
[26] Torres C, Vitalis E A, Baker B R, et al. LAVA:An open-source approach to designing LAMP (loop-mediated isothermal amplification) DNA signatures[J].BMC Bioinformatics, 2011, 12(1):1-7.
[27] Chen H T, Zhang J, Yang S H, et al. Rapid detection of porcine parvovirus DNA by sensitive loop-mediated isothermal amplification[J]. Journal of Virological Methods, 2009, 158(1-2):100-103.
[28] Aebischer A, Wernike K, Hoffmann B, et al. Rapid genome detection of Schmallenberg virus and bovine viral diarrhea virus by use of isothermal amplification methods and high-speed Real-time reverse transcriptase PCR[J]. Journal of Clinical Microbiology, 2014, 52(6):1883-1892.
[29] Abd E W A, El-Deeb A, El-Tholoth M, et al. A portable reverse transcription recombinase polymerase amplification assay for rapid detection of foot-and-mouth disease virus[J]. PLoS One, 2013, 8(8):1-10.
[30] Wang J, Wang J, Liu L, et al. Rapid detection of porcine circovirus 2 by recombinase polymerase amplification[J]. Journal of Veterinary Diagnostic Investigation, 2016, 28(5):574-578.

Development of the Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Parvovirus

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	XU Yunming, WEI Libin, ZHOU Yanyang, LI Yehui, SUN Zhiyuan, ZHONG Siyuan, CHEN Aining, GUO Lijuan, LIU Zengshan, REN Honglin. Establishment of Loop-mediated Isothermal Amplification (LAMP) Method for Klebsiella pneumoniae [J]. , 2018, 45(11): 3003-3010.
[2]	LIU Wenjun, HUANG Yunmao, YANG Youtian, XU Danning, CAO Nan, ZHOU Derong, TIAN Yunbo. Establishment of RT-LAMP Visual Detection Method for Goose Newcastle Disease Virus [J]. , 2018, 45(1): 22-31.
[3]	LI Dan-ni, HUANG Hua, ZHANG Jing, WANG Xue-sheng, GU Yi-wen. Preparation and Properties Analysis of Three β₂-agonists Fluorescent Immunochromatographic Strip [J]. , 2017, 44(8): 2437-2442.
[4]	CHEN Xi-wen, WANG Qian, YIN Miao, LI Lian, LUO Wen-tao, YANG Feng, GUO Ai-wei, ZHOU Jie-long, WANG Xiong-qing. Establishment of RT-LAMP for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus [J]. , 2017, 44(6): 1630-1636.
[5]	HAN Xiao-ya, CHEN Dong-mei, WANG Yu-lian, TAO Yan-fei, DONG Guo-liang, PENG DA-peng, YUAN Zong-hui. Research Progress on Antibodies Rapid Detection Technology for Veterinary Synthetic Antibacterial Agents [J]. , 2017, 44(6): 1869-1876.
[6]	WANG Xin, SHI Da, DONG Hui, CAO Li-yan, CHEN Jian-fei, SHI Hong-yan, ZHANG Xin, FENG Li. Construction and Characterization of the Yeast Two-hybrid cDNA Library of ST Cells [J]. , 2017, 44(3): 667-672.
[7]	LUO Ya-kun, LIANG Lin, WANG Jing, LIU Cun, LIU Qi, LIU Chang, LIN Wen-cheng, CUI Shang-jin. Establishment and Application of Loop-mediated Isothermal Amplification for Rapid Detection of Porcine Epidemic Diarrhea Virus [J]. , 2017, 44(2): 344-349.
[8]	LV Ji-zhou, FAN Yan-ru, FENG Chun-yan, YUAN Xiang-fen, WU Shao-qiang. Establishment of Real-time Recombinase Polymerase Amplification for Detection of Porcine Epidemic Diarrhoea Virus [J]. , 2017, 44(12): 3434-3439.
[9]	HADENG Chu-riya, FAN Xiao-xu, ZHAO Yong-gang, WANG Shu-juan, ZHANG Zhi-cheng, GE Sheng-qiang, LI Lin, WU Xiao-dong, WANG Zhi-liang. Establishment of a Real-time Fluorescent Recombinase Polymerase Amplification (RPA) for the Detection of African Swine Fever Virus [J]. , 2017, 44(11): 3270-3277.
[10]	WU Zhi, XIA Wen-long, CAI Shu-dong, GUO Chang-ming, YUAN Wei-feng, WANG Yong-juan. Establishment and Application of Visual RT-LAMP for Rapid Detection of Duck Tembusu Virus [J]. , 2017, 44(11): 3334-3339.
[11]	BAI Yu, ZHANG Jing, HU Jing-yan, XU Yong-peng, WANG Shu-ming. Study on Colloidal Gold Immunochromatographic Assay for Rapid Detection of Estradiol in Milk [J]. , 2017, 44(11): 3351-3357.
[12]	NI Teng-teng, PENG Da-peng, XIE Shu-yu, CHEN Dong-mei, WANG Yu-lian, PAN Yuan-hu, TAO Yan-fei, YUAN Zong-hui. Research Progress on Rapid Determination Technology of Avermectins in Feeds and Animal Products [J]. , 2016, 43(9): 2344-2351.
[13]	WU Yu-xi, LI Pu-jun, WANG Juan, YANG Fa-long. Loop-mediated Isothermal Amplification Method and Its Application in Pathogen Detection [J]. , 2016, 43(2): 389-393.
[14]	WANG Cai-xia, FENG Chun-yan, WANG Qiao-li, YUAN Xiang-fen, DENG Jun-hua, LV Ji-zhou, WU Shao-qiang. Establishment of Duplex Loop-mediated Isothermal Amplification Method for Simultaneous Detection of Shellfish Infected with Perkinsus and Bonamia [J]. , 2015, 42(8): 1935-1942.
[15]	ZHANG Li, LI Kai, TIAN Guang-ling, LI De-feng, WANG Wei-song, HE Hou-jun. Development of Loop-mediated Isothermal Amplification Assay for Detection of Torque teno sus Virus Type 2 [J]. , 2015, 42(5): 1098-1103.